Molecular cloning and thermal stress-induced expression of a pi-class glutathione S-transferase (GST) in the Antarctic bivalve Laternula elliptica

被引:48
作者
Kim, Meesun [1 ]
Ahn, In-Young [1 ]
Cheon, Jina [1 ]
Park, Hyun [1 ]
机构
[1] Korea Polar Res Inst, KORDI, Inchon 406840, South Korea
来源
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY | 2009年 / 152卷 / 02期
关键词
Antarctic; Laternula elliptica; Pi class glutathione S-transferases; Recombinant GST; Thermal stress; OXIDATIVE STRESS; MYTILUS-GALLOPROVINCIALIS; IDENTIFICATION; TEMPERATURE; RAT; PROTEINS; KINASE; THETA; GENE; P1-1;
D O I
10.1016/j.cbpa.2008.09.028
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Glutathione S-transferases (GSTs) are multifunctional phase II detoxification enzymes that catalyze the attachment of electrophilic substrates to glutathione. The pi-class GST cDNA (leGSTp) was cloned from the cold-adapted Antarctic bivalve Laternula elliptica. We used degenerated primers designed based on highly conserved regions of known mollusk GSTs to amplify the corresponding L. elliptica mRNA. Full-length cDNA was obtained by rapid amplification of cDNA ends (RACE). The full sequence of the GST cDNA was 1189 bp in length, with a 5' untranslated region (UTR) of 74 bp, a 3' UTR of 485 bp, and an open reading frame of 630 bp encoding 209 amino acid residues with an estimated molecular mass of 23.9 kDa and an estimated isoelectric point of 8.3. Quantitative RT-PCR confirmed basal expression of leGSTp, which was up-regulated upon heat treatment (10 degrees C for different time periods) by a factor of 2.3 (at 24 h) and 2.7 (at 48 h) in the digestive gland and gill tissues, respectively. The recombinant leGSTp expressed in Escherichia coli was purified by affinity chromatography and characterized. The purified leGSTp exhibited high activity towards the substrates ethacrynic acid (ECA) and 1-chloro-2,4-dinitrobenzene (CDNB). The recombinant leGSTp had a maximum activity at approximately pH 8.0, and its optimum temperature was 35 degrees C. (C) 2008 Elsevier Inc. All rights reserved.
引用
收藏
页码:207 / 213
页数:7
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